Figure 3
From: Quantifying cell death induced by doxorubicin, hyperthermia or HIFU ablation with flow cytometry
Flow cytometric studies (dot plot, histograms) of viable HT29 cell suspension exposed to high concentration doxorubicin (RIGHT) 2 h previously or not (LEFT). Samples are either unstained detected using FL2 channel (excitation/emission 488/585 ± 42 nm) (top row), stained with propidium iodide detected using the same FL2 channel (middle row), or the far-red L/D stain detected using FL-4 channel (633/660 ± 16 nm) (bottom row). On the logarithmic FL-4 scale, the vertical line represents the fluorescence gating threshold, with events to the left of the line representing viable cells, and events to the right of the line representing positive staining by the viability, i.e. what might be expected to be non-viable cells. Percentage of cell viability (top left, dot plot) vs. death (top right, dot plot) are demonstrated within the gated areas. However, note that the PI stain gives a misleading impression of near complete cell death in the only very recently doxorubicin-exposed cells due to the overlap with doxorubicin’s wide fluorescent emission profile, when in reality there has been insufficient time for true cell necrosis. Events with FSC < 100 discarded, presumed to represent debris and/or impurities, with around 90% events included for analysis. Plots generated using FlowJo Data Analysis Software.
